Objective To investigate the role of C-Jun N-terminal kinase (JNK) in epithelial mesenchymal transition (EMT) induced by transforming growth factor β1 (TGF-β1) in rat peritoneal mesothelial cells(RPMCs). Methods RPMCs were harvested from the peritoneum of male Sprague-Dawley rats, then cultured in DMEM/F12 medium with 15% (V/V) FBS. After stimulation with TGF-β1, the expression of a-smooth muscle actin (α-SMA), E-cadherin and collagen I were detected in RPMCs. In some groups, the ceils were pretreated with SP600125, a specific inhibitor of JNK, for 4 hours before incubation with TGF-β1. The protein expression of phosphorylated JNK was detected by Western blotting. The mRNA and protein expression ofα-SMA, E-cadherin and collagen I were examined with RT-PCR and Western blotting, respectively.The intracellular distribution and expression of α-SMA was determined by indirect immunofluorescence. Results TGF-β1 could significantly increase the expression of α-SMA and collagen I, and decrease the expression of E-cadherin in RPMCs. TGF-α1 could stimulate the expression of phosphorylated JNK at 5 minutes with the peak at 10 minutes (P<0.01). The addition of SP600125 effectively inhibited TGF-β1-induced high expression of α-SMA and collagen I (P<0.05), and prevented TGF-β1-induced down-regulation of E-cadherin expression in RPMCs (P<0.05). The indirect immunofluorescence showed that the expression of intracellular α-SMA in RPMCs stimulated by TGF-β1 for 48 h increased significantly, which could be inhibited by SP600125. Conclusions JNK regulates epithelial mesenchymal transition induced by TGF-β1 in rat peritoneal mesothelial cells. JNK inhibitor may be used as a novel therapeutic agent for peritoneal fibrosis.

Objective To explore the brain biochemical changes in the frontal lobe of adolescents with depression using proton magnetic resonance spectroscopy (1H-MRS). Methods Twenty-four patients and twenty-three healthy subjects matched for age, sex and education level were enrolled in the study. All the subjects underwent multivoxel 1H MRS to measure the bilateral metabolic levels of N-acetylaspartate (NAA), choline (Cho), creatine (Cr) in the prefrontal lobes. Results The NAA/Cr and Cho/Cr ratios in the left dorsolateral prefrontal white matter of the depressive adolescents were significantly lower than those of the healthy subjects [NAA/Cr: 1.67 ± 0.32, t = 3.126, P = 0.004; Cho/Cr: 1.28 ± 0.30, t = 2.362, P = 0.024], and the ratios of NAA/Cr in the right dorsolateral prefrontal white matter of the depressive adolescents was also significantly lower than that of the healthy subjects [NAA/Cr:1.65 ± 0.26, t=2.969, P=0.006]. There was no significant difference in the metabolic ratios in the bilateral anterior cingulate gray matter between the depressive adolescents and the healthy controls. Conclusions Biochemical abnormalities in prefrontal white matter are involved in the pathophysiology of depression. Importantly , these abnormalities are already present early in the course of the disorder.

OBJECTIVETo analyze medication laws of Chinese medicine (CM) treatment in hypertension patients with yin deficiency yang hyperactivity syndrome.

METHODSChina National Knowledge Infrastructure (CNKI, Jan. 1979-Dec 2014), Chinese Scientific Journals Database (VIP, Jan 1989-Dec2014), Chinese Biomedical Literature Database (CBM, Jan.1978-Dec.2014), Wanfang Database (Jan 1990-Dec 2014) were retrieved by using "hypertension", "CM", "Chinese herbs", "syndrome" as keywords. Totally 149 literatures concerning CM treatment for hypertension patients with yin deficiency yanghyperactivity syndrome were included in this study. The herbs database was established by SPSS20.0,and correlation laws were analyzed by SAS9.3. With the Pajek3.1, results were presented visually withcomplex networks.

RESULTSThere were 149 literatures including 131 kinds of herbs with 1,598 frequencies. The conventional compatibility program of herbs for asthenic yin and predominant yang syndrome of hypertension were two toothed achyranthes root, tall gastrodia rhizome, Cassia obtusifolia L., eucommiabark, baikal skullcap root, and so on, about 29 kinds. Of them, core herbs were two toothed achyranthes root, tall gastrodia rhizome, Cassia obtusifolia L., poria, prepared rhizome of rehmannia, oriental water-plantain tuber, asiatic cornelian cherry fruit, Uncariae Rhynchophylla, common yam rhizome, the rootbark of the peony tree, and so on.

CONCLUSIONMedication laws of CM treatment in hypertension patientswith yin deficiency yang hyperactivity syndrome obtained by analysis of complex networks reflected thetherapeutics of nourishing yin to suppress yang, which could further provide reference for clinical studies.

Asian Continental Ancestry Group ; China ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Hypertension ; therapy ; Medicine, Chinese Traditional ; Yin Deficiency ; therapy ; Yin-Yang

Objective To investigate the effect of angiotensinⅡ(AngⅡ) type 1 receptor blocker losartan on the cyclooxygenase 2 (COX-2) expression in metabolic syndrome (MS)kidney. Methods Seven-week-old male obese Zucker rats,a model of MS,were randomly divided into losartan treated and untreated group,and lean Zucker rats were used as controls.The obese Zucker rata of treated group received losartan for 4 months continuously.COX-2 expression was examined for all rats after 4 months.AngⅡ-stimulated mesangial cells and cortical tissue from AngⅡ-infused C57BL/6 mouse kidney by osmotic minipumps were used in this study.RNA and protein were obtained from renal cortical tissue or mesangial cells for RT-PCR and Western blot.Results Compared to the lean controls,obese Zucker rats showed a significant increase of COX-2 expression in the renal cortical tissue and these abnormalities were prevented by administration of losartan. Furthermore,the direct stimulation of AngⅡincreased COX-2 expression in mesangial cells in vitro and renal cortical tissue in vivo.Conclusions MS-induced COX-2 expression in the kidney is regulated by AngⅡ.Losartan as a non COX-2 inhibitor can protect MS kidney,at least in part,by inhibition of COX-2 activation.

To compare the differences of the active ingredient contents and the sulfur dioxide residue in Astragali Radix before and after sulfur fumigation and provide a basis for establishing an alternative processing method. Astragali Radix, harvested at the same time in Longxi Gansu, were processed with different methods. high performance liquid chromatography (HPLC) was used to determine the contents of the active ingredients in Astragali Radix and the revised method of the pharmacopoeia of China in 2011 was applied to determine the sulfur dioxide residue. The results show that the three-fold sulfur-fumigation group has the highest level of astragaloside IV and the dried sulfur-fumigation group with 10% water has the lowest level; the content of calycosin-7-O-β-D-glucoside is the highest in naturally dried group and the lowest in the group of sulfur fumigating for 3 times; the sulfur dioxide residue of all sulfur-fumigation groups exceeds certain limit significantly and the group of sulfur fumigating for 3 times reaches the highest level.
Astragalus Plant ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Fumigation ; adverse effects ; Sulfur Dioxide ; analysis ; Technology, Pharmaceutical ; methods

The contents of adenosine, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, parishin and sulfur dioxide residue were compared in differently-processed Gastrodiae Rhizoma to provide the basis for a reasonable processing method of Gastrodiae Rhizoma. The analysis was performed on a Merck Purospher STAR column (4.6 mm x 250 mm, 5 μm) with a mobile phase consisting of methanol and water (containing 0.1% formic acid) under gradient elution at a flow rate of 1.0 mL x min(-1). The eluates were detected at 270 nm, and the column temperature was 35°C. The content of adenosin, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxy-benzaldehyde and parishin in processing of boiling or sulfur-fumigated were lower than that of in processing of steaming. Furthermore, the sulfur dioxide residue of sulphur-fumigated groups exceed 400 mg x kg(-1). This stable and reliable method will contribute to the quality control of different processed Gastrodiae Rhizoma.
Drug Contamination ; Drugs, Chinese Herbal ; chemistry ; Gastrodia ; chemistry ; Sulfur Dioxide ; analysis ; Technology, Pharmaceutical ; methods

AIM: To investigate the expression of connective tissue growth factor (CTGF) and α-SMA in human lens epithelium cell (HLEC) line B3 after transfection by liposome-coated siRNA targeting CTGF.METHODS: HLECs were transfected with small interfering RNA (siRNA) targeting CTGF,labeled with 5`-fluorescein isothiocyanate (5`-FITC) and coated with lipofectamine.The transfection ratio was evaluated via fluorescence intensity.Cell counting kit-8 (CCK-8) assay was performed to assess cytoviability of both non-transfected and transfected HLECs.Quantitative RT-PCR,cell immunochemistry and Western blot analysis were conducted to detect the expression changes of CTGF and α-SMA after transfection.RESULTS: A highly effective transfection ratio was observed in siRNA co-transfected with lipofectamine.The transfection ratio reached 95% at 24h.The proliferation of HLECs was inhibited by siRNA after 72h transfection.The expression of CTGF and α-SMA significantly decreased in HLECs after transfected by CTGF siRNA for 24h.This effect was not found in negative control siRNA.CONCLUSIONS: SiRNA targeting CTGF decreased CTGF and α-SMA expression in HLECs,which is a potential therapeutic strategy for posterior capsular opacification.

Small intestinal obstruction is a common complication of primary gastrointestinal cancer or metastatic cancers.Patients with this condition are often poor candidates for surgical bypasses,and placement of self-expanding metal stent (SEMS) can be technically challenging.In this study,we examined the feasibility of combined application of single-balloon enteroscope (SBE) and colonoscope for SEMS placement in patients with malignant small intestinal obstruction.Thirty-four patients were enrolled in this study,among which 22 patients received SEMS placement by using SBE and colonoscope,while the other 12 patients received conservative medical treatment.The patients were followed up for one year.Stent placernent was technically feasible in 95.5％ (21/22).Clinical improvement was achieved in 86.4％ (19/22).For the 19 clinical success cases,the average time of benefits from a gastric outlet obstruction scoring system (GOOSS) increase ≥1 was 111.9±89.5 days.For the 12 patients receiving conservative medical treatment,no significant improvement in GOOSS score was observed.Moreover,a significant increase of Short-Form-36 health survey score was observed in the 19 patients at time of 30 days after stent placement.By Kaplan-Meier analysis,a significant survival improvement was observed in patients with successful SEMS placement,compared with patients receiving conservative medical treatment.Taken together,combined use of SBE and colonoscope makes endoscopic stent placement feasible in patients with malignant small intestinal obstruction,and patients can benefit from it in terms of prolonged survival and improved quality of life.

BACKGROUNDThe global outbreak of influenza A (H1N1) has led to the Ministry of Health of China listing it as one of the A-class infectious diseases. Pneumonia is the most serious complication of influenza A, commonly causing death. Populations are ordinarily susceptible to influenza A. This study aimed to investigate the imaging manifestation features of critical influenza A (H1N1) pneumonia and to improve its diagnostic techniques.

METHODSA total of seven death cases from critical influenza A (H1N1) pneumonia were retrospectively analyzed on their imaging manifestations and autopsy data. Pulmonary CT scanning was performed for five cases, with one receiving additional chest X-ray and chest CT scanning, and chest postero-anterior position X-ray examination was performed for other two. Autopsy was performed for five cases and postmortem examinations were performed for other two cases.

RESULTSThe seven cases of influenza A showed critical manifestations in 4 - 7 days after symptoms onset, with two having basic diseases of diabetes and one being pregnant. Extensive blurry high-density shadows of bilateral lungs were found in three cases, which were most obvious in middle and inferior parts of lungs. Pulmonary CT scanning revealed bilateral flaky parenchymal shadows in peripheral, dorsal and fundus segments of the middle-inferior parts of lungs, with one case of complicated pneumothorax, atelectasis and pleural effusion and another case of thin-walled cavity and dilated bronchi shadows in the superior parts of lungs.

CONCLUSIONSDiagnostic imaging is an important assessing tool for critical influenza A (H1N1) pneumonia. The imaging manifestations are characteristic instead of being specific. The definitive diagnosis can be made in combination with clinical examinations and laboratory tests.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; pathogenicity ; Influenza, Human ; diagnosis ; diagnostic imaging ; virology ; Male ; Middle Aged ; Pregnancy ; Radiography ; Retrospective Studies ; Young Adult

The purpose of this study was to evaluate whether the DNA vaccine containing idiotypic gene fragment of human B-cell lymphoma cell line Namalwa could elicit the specific anti-idiotypic immune response in vivo. The candidate gene fragment of the lymphoma cell, variable region of heavy chain (VH) of the membranous immunoglobulin, was amplified using Ig superfamily primers by means of RT-PCR. Also, the intact cDNA of murine monocyte chemoattractant protein (MCP-3) was cloned and used as the adjuvant molecular. The two gene fragments of VH and MCP-3 were fused together by 8aa linker peptide with recombinant PCR. Subsequently, the fusion gene fragment was cloned into eukaryonic expression vector pcDNA3.1 to construct DNA vaccine plasmid. Prior to the immunization, the transient transfection coupled with RT-PCR was performed to prove that the recombinant plasmid could express in eukaryonic cells in right way. Then two groups of mice were immunized by intramuscular injection with DNA vaccine and mock plasmid pcDNA3.1 respectively. Three times of injection were performed with 100 micro g plasmid respectively at the beginning of the experiment and 2, 4 weeks after the first injection for all the groups. FACS analysis was chosen to detect the antibodies recognizing lymphoma cells at different time following vaccination. The results demonstrated that specific anti-idiotypic antibody could be detected in the group of DNA vaccine immunized mice as early as eight weeks after the first immunization. Further test demonstrated that the anti-idiotypic antibody could maintain for at least twenty weeks with high titer. Anti-idiotypic antibodies were elicited in three of five mice of the DNA vaccine immunized group. The Abs of DNA vaccine immunized mice could only recognize Namalwa cell line instead of another unrelated human cell line A549. There is no cellular response detected in the DNA vaccine immunized mice. It is concluded that the DNA vaccine containing fused MCP3-VH sequence could elicit specific anti-idiotypic antibody against B-cell lymphoma in vivo and could be used in further study of DNA vaccine against B-cell lymphoma. The results would provide the basis for further studies and optimization of this therapeutic strategy on patients with B-lymphoproliferative disease.
Animals ; Antibodies, Anti-Idiotypic ; blood ; COS Cells ; Cancer Vaccines ; immunology ; Chemokine CCL7 ; Cytokines ; Humans ; Immunization ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Lymphoma, B-Cell ; immunology ; Mice ; Mice, Inbred BALB C ; Monocyte Chemoattractant Proteins ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, DNA ; immunology